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RU2010140227A - POLYEPEPTIDE POSSESSING THE ACTIVITY OF GLIOXALASE III, ENCODING ITS POLINUCLEOTID AND THEIR APPLICATION - Google Patents

POLYEPEPTIDE POSSESSING THE ACTIVITY OF GLIOXALASE III, ENCODING ITS POLINUCLEOTID AND THEIR APPLICATION Download PDF

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RU2010140227A
RU2010140227A RU2010140227/10A RU2010140227A RU2010140227A RU 2010140227 A RU2010140227 A RU 2010140227A RU 2010140227/10 A RU2010140227/10 A RU 2010140227/10A RU 2010140227 A RU2010140227 A RU 2010140227A RU 2010140227 A RU2010140227 A RU 2010140227A
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microorganism according
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Франсуа ВЁЛКЕ (FR)
Франсуа ВЁЛКЕ
Филипп СУКЕЛЬ (FR)
Филипп СУКЕЛЬ
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Метаболик Эксплорер (Fr)
Метаболик Эксплорер
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    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/56—Lactic acid
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0008—Oxidoreductases (1.) acting on the aldehyde or oxo group of donors (1.2)
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    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/18—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic polyhydric
    • C—CHEMISTRY; METALLURGY
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    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/24—Preparation of oxygen-containing organic compounds containing a carbonyl group
    • C12P7/26—Ketones

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Abstract

1. Модифицированный микроорганизм, обладающий модулированной ферментативной активностью глиоксалазы III, где эта активность полипептида, обладающего по меньшей мере 70%-ной идентичностью с последовательностью SEQ ID NO 1, аттенуирована или усилена. ! 2. Микроорганизм по п.1 с аттенуированной ферментативной активностью глиоксалазы III, где экспрессия нативного гена, кодирующего полипептид, обладающий по меньшей мере 70%-ной идентичностью с последовательностью SEQ ID NO 1, аттенуирована. ! 3. Микроорганизм по п.2, который дополнительно модифицирован для усиления продуцирования 1,2-пропандиола и/или ацетола. ! 4. Микроорганизм по п.3, который содержит по меньшей мере одну из приведенных ниже модификаций для усиления продуцирования 1,2-пропандиола и их комбинаций: ! - аттенуацию экспрессии по меньшей мере одного из следующих генов: ptsG, ptsH ptsl, crr, edd, eda, gloA, aldA, aldB, adhA, pflA, pflB, adhE, tpiA, gapA, pykA, pykF, ackA, pta, poxB, arcA и ndh. ! - усиление экспрессии по меньшей мере одного из следующих генов: galP, glk, ppsA, mgsA, yghD, yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC, gldA, fucO. ! - модификацию гена lpD, так что он имеет точечную мутацию, приводящую к замене аланина 55 на валин в белке, кодируемом этим геном. ! 5. Микроорганизм по п.3, который содержит по меньшей мере одну из приведенных ниже модификаций для усиления продуцирования ацетола и их комбинаций: ! - аттенуацию экспрессии по меньшей мере одного из следующих генов: ptsG, ptsH, ptsl, crr, edd, eda, gloA, aldA, aldB, ldhA, pflA, pflB, adhE, fpiA, gapA, pykA, pykF, ackA, pta, poxB и gldA. ! - усиление экспрессии по меньшей мере одного из следующих генов: galP, glk, ppsA, mgsA, yqhD, yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC. ! 6. Микроорганизм по п.1 с усиленной ферментативной активностью глиоксалазы III, где полинуклеотид, � 1. A modified microorganism having modulated glyoxalase III enzymatic activity, wherein the activity of a polypeptide having at least 70% identity with the sequence of SEQ ID NO 1 is attenuated or enhanced. ! 2. A microorganism according to claim 1 with attenuated glyoxalase III enzymatic activity, wherein the expression of a native gene encoding a polypeptide having at least 70% identity with the sequence of SEQ ID NO 1 is attenuated. ! 3. The microorganism according to claim 2, which is further modified to enhance the production of 1,2-propanediol and/or acetol. ! 4. The microorganism according to claim 3, which contains at least one of the following modifications to enhance the production of 1,2-propanediol and combinations thereof: ! - attenuation of the expression of at least one of the following genes: ptsG, ptsH ptsl, crr, edd, eda, gloA, aldA, aldB, adhA, pflA, pflB, adhE, tpiA, gapA, pykA, pykF, ackA, pta, poxB, arcA and ndh. ! - increased expression of at least one of the following genes: galP, glk, ppsA, mgsA, yghD, yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC, gldA, fucO. ! - modification of the lpD gene so that it has a point mutation resulting in the replacement of alanine 55 by valine in the protein encoded by this gene. ! 5. The microorganism according to claim 3, which contains at least one of the following modifications to enhance the production of acetol and combinations thereof: ! - attenuation of the expression of at least one of the following genes: ptsG, ptsH, ptsl, crr, edd, eda, gloA, aldA, aldB, ldhA, pflA, pflB, adhE, fpiA, gapA, pykA, pykF, ackA, pta, poxB and gldA. ! - increased expression of at least one of the following genes: galP, glk, ppsA, mgsA, yqhD, yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC. ! 6. The microorganism according to claim 1 with enhanced enzymatic activity of glyoxalase III, where the polynucleotide, �

Claims (15)

1. Модифицированный микроорганизм, обладающий модулированной ферментативной активностью глиоксалазы III, где эта активность полипептида, обладающего по меньшей мере 70%-ной идентичностью с последовательностью SEQ ID NO 1, аттенуирована или усилена.1. A modified microorganism having modulated enzymatic activity of glyoxalase III, where this activity of a polypeptide having at least 70% identity with the sequence of SEQ ID NO 1 is attenuated or enhanced. 2. Микроорганизм по п.1 с аттенуированной ферментативной активностью глиоксалазы III, где экспрессия нативного гена, кодирующего полипептид, обладающий по меньшей мере 70%-ной идентичностью с последовательностью SEQ ID NO 1, аттенуирована.2. The microorganism according to claim 1 with attenuated enzymatic activity of glyoxalase III, where the expression of a native gene encoding a polypeptide having at least 70% identity with the sequence of SEQ ID NO 1 is attenuated. 3. Микроорганизм по п.2, который дополнительно модифицирован для усиления продуцирования 1,2-пропандиола и/или ацетола.3. The microorganism according to claim 2, which is further modified to enhance the production of 1,2-propanediol and / or acetol. 4. Микроорганизм по п.3, который содержит по меньшей мере одну из приведенных ниже модификаций для усиления продуцирования 1,2-пропандиола и их комбинаций:4. The microorganism according to claim 3, which contains at least one of the following modifications to enhance the production of 1,2-propanediol and their combinations: - аттенуацию экспрессии по меньшей мере одного из следующих генов: ptsG, ptsH ptsl, crr, edd, eda, gloA, aldA, aldB, adhA, pflA, pflB, adhE, tpiA, gapA, pykA, pykF, ackA, pta, poxB, arcA и ndh.attenuation of expression of at least one of the following genes: ptsG, ptsH ptsl, crr, edd, eda, gloA, aldA, aldB, adhA, pflA, pflB, adhE, tpiA, gapA, pykA, pykF, ackA, pta, poxB, arcA and ndh. - усиление экспрессии по меньшей мере одного из следующих генов: galP, glk, ppsA, mgsA, yghD, yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC, gldA, fucO.- enhancing the expression of at least one of the following genes: galP, glk, ppsA, mgsA, yghD, yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC, gldA, fucO. - модификацию гена lpD, так что он имеет точечную мутацию, приводящую к замене аланина 55 на валин в белке, кодируемом этим геном.- modification of the lpD gene, so that it has a point mutation, leading to the replacement of alanine 55 with valine in the protein encoded by this gene. 5. Микроорганизм по п.3, который содержит по меньшей мере одну из приведенных ниже модификаций для усиления продуцирования ацетола и их комбинаций:5. The microorganism according to claim 3, which contains at least one of the following modifications to enhance the production of acetol and their combinations: - аттенуацию экспрессии по меньшей мере одного из следующих генов: ptsG, ptsH, ptsl, crr, edd, eda, gloA, aldA, aldB, ldhA, pflA, pflB, adhE, fpiA, gapA, pykA, pykF, ackA, pta, poxB и gldA.attenuation of expression of at least one of the following genes: ptsG, ptsH, ptsl, crr, edd, eda, gloA, aldA, aldB, ldhA, pflA, pflB, adhE, fpiA, gapA, pykA, pykF, ackA, pta, poxB and gldA. - усиление экспрессии по меньшей мере одного из следующих генов: galP, glk, ppsA, mgsA, yqhD, yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC.- enhancing the expression of at least one of the following genes: galP, glk, ppsA, mgsA, yqhD, yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC. 6. Микроорганизм по п.1 с усиленной ферментативной активностью глиоксалазы III, где полинуклеотид, содержащий последовательность SEQ ID NO 2, претерпевает гиперэкспрессию.6. The microorganism according to claim 1 with enhanced enzymatic activity of glyoxalase III, where a polynucleotide containing the sequence of SEQ ID NO 2 undergoes overexpression. 7. Микроорганизм по п.6, который дополнительно модифицирован для усиления продуцирования лактата.7. The microorganism according to claim 6, which is further modified to enhance the production of lactate. 8. Микроорганизм по п.7, который содержит по меньшей мере одну из приведенных ниже модификаций для усиления продуцирования лактата и их комбинаций:8. The microorganism according to claim 7, which contains at least one of the following modifications to enhance the production of lactate and their combinations: - аттенуацию экспрессии по меньшей мере одного из следующих генов: ptsG, ptsH, ptsl, crr, edd, eda, gloA, aldA, aldB, pflA, pflB, frdABCD, adhE, gapA, pykA, pykF, ackA, pta, poxB, yqhD, yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC, gldA, lldD, dld и tpiA.- attenuation of expression of at least one of the following genes: ptsG, ptsH, ptsl, crr, edd, eda, gloA, aldA, aldB, pflA, pflB, frdABCD, adhE, gapA, pykA, pykF, ackA, pta, poxB, yqhD , yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC, gldA, lldD, dld and tpiA. - усиление экспрессии по меньшей мере одного из следующих генов: galP, glk, ppsA и mgsA.- enhancing the expression of at least one of the following genes: galP, glk, ppsA and mgsA. 9. Микроорганизм по любому из пп.1-8, выбранный из группы, состоящей из бактерий, дрожжей и грибов.9. The microorganism according to any one of claims 1 to 8, selected from the group consisting of bacteria, yeast and fungi. 10. Микроорганизм по п.9, где бактерии выбраны из группы, состоящей из Enterobacteriaceae, Bacillaceae, Streptomycetaceae и Corynebacteriaceae.10. The microorganism according to claim 9, where the bacteria are selected from the group consisting of Enterobacteriaceae, Bacillaceae, Streptomycetaceae and Corynebacteriaceae. 11. Микроорганизм по п.10, выбранный из группы, состоящей из Escherichia coli, Bacillus subtills, Clostridium acetobutylicum и Corynebacterium glutamicum.11. The microorganism of claim 10, selected from the group consisting of Escherichia coli, Bacillus subtills, Clostridium acetobutylicum and Corynebacterium glutamicum. 12. Способ модулирования ферментативной активности глиоксалазы III в микроорганизме, где активность полипептида, обладающего по меньшей мере 70%-ной идентичностью с последовательностью SEQ ID NO 1, усилена или аттенуирована в этом микроорганизме.12. A method of modulating the enzymatic activity of glyoxalase III in a microorganism, where the activity of a polypeptide having at least 70% identity with the sequence of SEQ ID NO 1 is enhanced or attenuated in this microorganism. 13. Способ получения 1,2-пропандиола и/или ацетола, где микроорганизм по любому из пп.2-5 выращивают в соответствующей культуральной среде, содержащей источник углерода, и выделяют 1,2-пропандиол и/или ацетол.13. A method of producing 1,2-propanediol and / or acetol, where the microorganism according to any one of claims 2 to 5 is grown in an appropriate culture medium containing a carbon source, and 1,2-propanediol and / or acetol is isolated. 14. Способ получения лактата, где микроорганизм по любому из пп.6-8 выращивают в соответствующей культуральной среде, содержащей источник углерода, и выделяют лактат.14. A method of producing lactate, where the microorganism according to any one of claims 6 to 8 is grown in an appropriate culture medium containing a carbon source, and lactate is isolated. 15. Способ по пп.13 или 14, где выделенный 1,2-пропандиол и/или ацетол или лактат очищают. 15. The method according to claims 13 or 14, wherein the isolated 1,2-propanediol and / or acetol or lactate is purified.
RU2010140227/10A 2008-03-18 2009-03-16 POLYEPEPTIDE POSSESSING THE ACTIVITY OF GLIOXALASE III, ENCODING ITS POLINUCLEOTID AND THEIR APPLICATION RU2010140227A (en)

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PCT/EP2008/053224 WO2009115114A1 (en) 2008-03-18 2008-03-18 Polypeptide having glyoxylase iii activity, polynucleotide encoding the same and uses thereof
EPPCT/EP2008/053224 2008-03-18

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CA2715737A1 (en) 2009-09-24
IL207648A0 (en) 2010-12-30
BRPI0909756A2 (en) 2015-08-25
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US20110014666A1 (en) 2011-01-20
JP2011515083A (en) 2011-05-19

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